1. Field Of The Invention
This invention relates to monoclonal antibodies produced by hybridoma cell lines created by the fusion of myeloma cell lines with lymphocytes.
The invention further relates to methods for detecting anaerobic micro-organisms in clinical samples; more specifically the present invention relates to methods for detecting Treponema species, potential etiological agents in periodontal disease (Moore, et al. Infection and Immunity, 38, 1137, (1982)).
It has now been found that the fusion of a mouse myeloma cell line, X63-Ag8.653, with a BALB/c mouse lymphocyte creates a hybridoma which produces a monoclonal antibody which is capable of specifically detecting at least eight strains of Treponema denticola.
2. Discussion Of The Prior Art
It is possible that certain spirochetes may be etiologic agents of severe periodontitis in adults or may serve as diagnostic indicators of this disease. Moore, et al. has described the isolation of ten Treponema species from plaque samples of patients with severe periodontitis. (Infection and Immunity, 38, 1137 (1982)). One of the treponemes more frequently isolated from diseased sites was identified as Treponema denticola. Treponema denticola and four other oral treponemes were also found to be among the most likely causative agents of moderate periodontitis. (Moore, et al., Infection and Immunity, 42, 510 (1983)). The role of specific spirochetes in periodontitis is unclear, since it is difficult and time-consuming to culture, purify, and positively identify Treponema denticola from clinical specimens. Therefore, immunochemical assays with specific monoclonal antibodies would greatly facilitate the detection and evaluation of this spirochete in human clinical studies.
Before the use of hybridomas to produce monoclonal antibodies, species specific antibodies were made by absorbing polyclonal animal sera with cross-reacting antigens. However, antisera prepared in animals are heterogeneous, unpredictable, and very limited in supply. The advent of a classical technique to produce monoclonal antibodies (Kohler and Milstein, Nature, 256, 495 (1975)) has allowed for the generation of immunodiagnostic reagents which are highly specific, homogeneous, and unlimited in supply.
A simplified technique based on the Kohler and Milstein methodology for creating hybridomas that produce monoclonal antibodies was described by Fazekas de St. Groth and Scheidegger (Journal of Immunological Methods, 35, 1 (1980)). The conditions described by Fazekas de St. Groth and Scheidegger have been utilized frequently; these conditions were modified to obtain a hybridoma producing a monoclonal antibody specific for fourteen strains of Bacteroides gingivalis (Simonson, et al., Journal of Dental Research, 65, 95 (1986)).
In addition, these conditions were again modified to produce the monoclonal antibody of the present invention (Simonson, et al., Infection and Immunity, 56,, 60 (1988)).
Recently, Strosberg, et al. disclosed in U.S. Pat. No. 4,780,407 a process for the immunological determination of Legionella pneumophilia bacteria using a monoclonal antibody produced by a hybridoma cell-line.